Energy-activated targeted cancer therapy

ABSTRACT

The present invention is drawn to methods and systems for administering a therapy to a target tissue or target composition in a mammalian subject, using an ultrasonic energy source that preferably transmits energy to a treatment site transcutaneously. The method provides for administering to the subject a therapeutically effective amount of a targeted substance, which preferably selectively binds to the target tissue. Energy at a wavelength or waveband corresponding to that which is absorbed by the targeted substance is then administered. The energy intensity is relatively low, but a high total fluence is employed to ensure the activation of the targeted energy-activated agent or targeted prodrug product. The claimed energy-activated targeted therapy is useful in the treatment of specifically selected target tissues, such as vascular endothelial tissue, the abnormal vascular walls of tumors, solid tumors of the head and neck, tumors of the gastrointestinal tract, tumors of the liver, tumors of the breast, tumors of the prostate, tumors of the lung, nonsolid tumors, malignant cells of the hematopoietic and lymphoid tissue and other lesions in the vascular system or bone marrow, and tissue or cells related to autoimmune and inflammatory disease.

RELATED APPLICATIONS

[0001] This application is a divisional application of copending U.S.patent application Ser. No. 09/271,575, filed Mar. 18, 1999, to JamesChen entitled “TARGETED TRANSCUTANEOUS CANCER THERAPY.”

[0002] Benefit of priority under 35 U.S.C. §119(e) to the followingprovisional applications is claimed herein: U.S. provisional applicationSerial No. 60/116,234 to James Chen, filed Jan. 15, 1999, entitled“TARGETED TRANSCUTANEOUS CANCER THERAPY.”

[0003] The above-noted applications and provisional applications areincorporated by reference in their entirety.

FIELD OF THE INVENTION

[0004] This invention generally relates to the delivery to a tumortarget site of a therapeutically effective amount of a photosensitizingagent that is activated by a relatively low fluence rate or level ofintensity of light administered over a prolonged period of time, andmore specifically, to the delivery of a photosensitizing agent that istargeted to bind with cancerous cells at the target site.

BACKGROUND OF THE INVENTION

[0005] One form of energy activated therapy for destroying abnormal ordiseased tissue is photodynamic therapy (PDT). PDT is a two-steptreatment process, which has received increasing interest as a mode oftreatment for a wide variety of different cancers and diseased tissue.The first step in this therapy is carried out by administering aphotosensitive compound systemically by ingestion or injection, ortopically applying the compound to a specific treatment site on apatient's body, followed by illumination of the treatment site withlight having a wavelength or waveband corresponding to a characteristicabsorption waveband of the photosensitizer. The light activates thephotosensitizing compound, causing singlet oxygen radicals and otherreactive species to be generated, leading to a number of biologicaleffects that destroy the abnormal or diseased tissue, which has absorbedthe photosensitizing compound. The depth and volume of the cytotoxiceffect on the abnormal tissue, such as a cancerous tumor, depend in parton the depth of the light penetration into the tissue; thephotosensitizer concentration and its cellular distribution, and theavailability of molecular oxygen which will depend upon the vasculaturesystem supplying the abnormal tissue or tumor.

[0006] Various types of PDT light sources and their methods of use havebeen described in the prior art literature. However, publicationsdescribing appropriate light sources and the effects of transcutaneouslight delivery to internal treatment sites within a patient's body, forPDT purposes, are relatively limited in number. It has generally beenaccepted that the ability of a light source external to the body tocause clinically useful cytotoxicity during PDT is limited in depth to arange of 1-2 cm or less, depending on the photosensitizer.

[0007] Treatment of superficial tumors in this manner has beenassociated with inadvertent skin damage due to accumulation of thephotosensitizer in normal skin tissue, which is a property of allsystemically administered photosensitizers in clinical use. For example,clinically useful porphyrins such as PHOTOPHRIN™ (a QLT, Ltd. brand ofsodium porfimer) are associated with general dermal photosensitivitylasting up to six weeks. PURLYTIN™ which is a brand of purpurin, andFOSCAN™, which is brand of chlorin, sensitize the skin to light for atleast several weeks, so that patients to whom these drugs areadministered must avoid exposure to sunlight or other bright lightsources during this time to avoid unintended phototoxic effects on thenormal dermal tissue. Indeed, efforts have been made to developphotoprotectants to reduced skin photosensitivity (see, for example:Dillon et al,, “Photochemistry and Photobiology,” 48(2): 235-238(1988);—and Sigdestad et al., British J. of Cancer, 74:S89-S92, (1996)).

[0008] Recently,it has been reported that a relatively intense externallaser light source might be employed transcutaneously to causetwo-photon absorption by a photosensitizer at a greater depth within apatient's body, so that it is theoretically possible to cause a verylimited volume of cytotoxicity in diseased tissue at greater depths thanpreviously believed possible. However, no clinical studies exist tosupport this contention. One would expect that the passage of an intensebeam of light through the skin would lead to the same risk of phototoxicinjury to non-target normal tissues, such as skin and subcutaneousnormal tissue, if this light is applied in conjunction with asystemically administered photosensitizer.

[0009] For example, one PDT modality discloses the use of an intenselaser source to activate a photosensitizer drug within a preciselydefined boundary (see U.S. Pat. No. 5,829,448, Fisher et al., “Methodfor improved selectivity in photo-activation of molecular agents”). Thetwo-photon methodology requires a high power, high intensity laser fordrug activation using a highly collimated beam, with a high degree ofspatial control. For a large tumor, this treatment is not practical,since the beam would have to be swept across the skin surface in somesort of set, repeating pattern, so that the beam encompasses the entirevolume of the tumor. Patient or organ movement would be a problem,because the beam could become misaligned. Exposure of normal tissue orskin in the path of the beam and subcutaneous tissue photosensitivity isnot addressed in the prior art literature.

[0010] Any photosensitizer absorbed by normal tissue in the path of thebeam will likely be activated and cause unwanted collateral normaltissue damage. Clearly, it would be preferable to employ a techniquethat minimizes the risk of damage to normal tissue and which does notdepend upon a high intensity laser light source to produce two photoneffects. Further, it would be preferable to provide a prolonged exposureof an internal treatment site with light at a lower fluence rate orlower intensity, which tends to reduce the risk of harm to non-targettissue or skin and subcutaneous normal tissue and reduces any collateraltissue damage due to phototoxicity.

[0011] Other PDT modalities have employed the use of a light sourceproducing a low total fluence delivered over a short time period toavoid harm to skin caused by activation of a photosensitizer and havetimed the administration of such drugs to better facilitate destructionof small tumors in animals (see, for example, U.S. Pat. No. 5,705,518,Richter et al.). However, although not taught or suggested by the priorart, it would be preferable to employ a light source that enables arelatively large total fluence PDT, but at a lower intensity so thatlarger tumor volumes can more readily be treated.

[0012] If, as is often the case, a target tumor tissue lies below anintact cutaneous layer of normal tissue, the main drawbacks of alltranscutaneous illumination methods, whether they be external laser orexternal non-laser light sources, are: (1) the risk of damage tonon-target tissues, such as the more superficial cutaneous andsubcutaneous tissues overlying the target tumor mass; (2) the limitedvolume of a tumor that can be treated; and (3) the limitation oftreatment depth. Damage to normal tissue lying between the light sourceand the target tissue in a tumor occurs due to the uptake ofphotosensitizer by the skin and other tissues overlying the tumor mass,and the resulting undesired photoactivation of the photosensitizerabsorbed by these tissues. The consequences of inadvertent skin damagecaused by transcutaneous light delivery to a subcutaneous tumor mayinclude severe pain, serious infection, and fistula formation. Thelimited volume of tumor that can be clinically treated and thelimitations of the light penetration below the skin surface in turn haveled those skilled in this art to conclude that clinical transcutaneousPDT is only suitable for treatment of superficial, thin lesions.

[0013] U.S. Pat. No. 5,445,608, Chen et al., discloses the use ofimplanted light sources for internally administering PDT. Typically, thetreatment of any internal cancerous lesions with PDT requires at least aminimally invasive procedure such as an endoscopic technique, forpositioning the light source proximate to the tumor, or open surgery toexpose the tumor site. There is some risk associated with any internalprocedure performed on the body. Clearly, there would be significantadvantage to a completely noninvasive form, of PDT directed tosubcutaneous and deep tumors, which avoids the inadvertent activation ofany photosensitizer in skin and intervening tissues. To date, thiscapability has not been clinically demonstrated nor realized. Only inanimal studies utilizing mice or other rodents with very thin cutaneoustissue layers, have very small superficial subcutaneous tumors beentreated with transcutaneously transmitted light. These minimal in vivostudies do not provide an enabling disclosure or even suggest howtranscutaneous light sources might safely be used to treat large tumorsin humans with PDT, however.

[0014] Another PDT modality in the prior art teaches the destruction ofabnormal cells that are circulating in the blood using light therapy,while leaving the blood vessels intact (see, for example: U.S. Pat. No.5,736,563, Richter et al.; WO 94/06424, Richter; WO 93/00005, Champan etal.; U.S. Pat. No. 5,484,803, Richter et al., and WO 93/24127, North etal. Instead, it might be preferable to deliberately damage and occludeblood vessels that form the vasculature supplying nutrients and oxygento a tumor mass, thus rendering a given volume of abnormal tissue in thetumor (not circulating cells) ischemic and anoxic and thus promoting thedeath of the tumor tissue serviced by these blood vessels.

[0015] To facilitate the selective destruction of the blood vessels thatservice a tumor, it would be desirable to selectively bind aphotosensitizing agent to specific target tissue antigens, such as thosefound on the epithelial cells comprising tumor blood vessels. Thistargeting scheme should decrease the amount of photosensitizing drugrequired for effective PDT, which in turn should reduce the total lightenergy, and the light intensity needed for effective photoactivation ofthe drug. Even if only a portion of a blood vessel is occluded as aresult of the PDT, downstream thrombosis is likely to occur, leading toa much greater volume of tumor necrosis compared to a direct cytotoxicmethod of destroying the tumor cells, in which the photosensitizer drugmust be delivered to all abnormal cells that are to be destroyed.

[0016] One method of ensuring highly specific uptake of aphotosensitizer by epithelial cells in tumor vessels would be to use theavidin-biotin targeting system. Highly specific binding of a targetedagent such as a PDT drug to tumor blood vessels (but not to the cells innormal blood vessels) is enabled by this two step system. While thereare reports in the scientific literature describing the binding betweenbiotin and streptavidin to target tumor cells, there are no reports ofusing this ligand-receptor binding pair to bind with cells in tumorvessels nor in conjunction with carrying out prolonged PDT lightexposure (see, for example: Savitsky et al., SPIE, 3191:343-353, (1997);and Ruebner et al., SPIE, 2625:328-332, (1996)). In a non-PDT modality,the biotin-streptavidin ligand-receptor binding pair has also beenreported as useful in binding tumor targeting conjugates withradionuclides (see U.S. Pat. No. 5,630,996, Reno et al.) and withmonoclonal antibodies (see Casalini et al.; J Nuclear Med,38(9):1378-1381, (1997)) and U.S. Pat. No. 5,482,698, Griffiths).

[0017] Other ligand-receptor binding pairs have been used in PDT fortargeting tumor antigens, but the prior art fails to teach their use inconjunction with targeting cells in blood vessels or treatment of large,established tumors (see, for example, Mew et al., J. of Immunol.,130(3): 1473-1477, (1983)).

[0018] High powered lasers are usually employed as a light source inadministering PDT to shorten the time required for the treatment (seeW.G. Fisher, et al., Photochemistry and Photobiology, 66(2):141-155,(1997)). However, it would likely be safer to use a low power,non-coherent light source that remains energized for two or more hoursto increase the depth of the photoactivation. This approach is contraryto the prior art that recommends PDT be carried out with a briefexposure from a high powered, collimated light source.

[0019] Recently, there has been much interest in the use ofanti-angiogenesis drugs for treating cancerous tumors by minimizing theblood supply that feeds a tumor's growth. However, targeting of tumorvessels using anti-angiogenesis drugs may lead to reduction in size ofsmall tumors and may prevent new tumor growth, but will likely beineffective in causing reliable regression of large, established tumorsin humans. However, by using a combination of anti-angiogenesis and aphotosensitizer in the targeting conjugate, it is likely that a largevolume tumor can be destroyed by administering PDT.

[0020] In treating large tumors, a staged procedure may be preferable inorder to control tumor swelling and the amount of necrotic tissueproduced as the PDT causes destruction of the tumor mass. For example,by activating a photosensitizer bound to tumor vessels in the center ofa large tumor and then sequentially expanding the treatment zone outwardin a stepwise manner, a large volume tumor can be gradually ablated in acontrolled fashion in order to prevent swelling due to edema andinflammation, which is problematic in organs such as the brain.

[0021] Delivered in vivo, PDT has been demonstrated to cause vesselthrombosis and vascular constriction, occlusion, and collapse. Andthough the treatment of very superficial, thin tumors has been reportedusing transcutaneous light, there are no clinical reports oftranscutaneous light activation being used to destroy deeper, thicktumors that are disposed more than 2 cm below the skin surface. Clearly,there is a need for a PDT paradigm that enables large volume tumors thatare disposed well below the surface of the skin to be destroyed withtranscutaneous light activation.

[0022] It is apparent that the usual method of administering PDT totreat bulky tumors, which relies on invasive introduction of opticalfibers, is not the best approach. It would be highly advantageous toapply light transcutaneously in a completely noninvasive method to treatsuch large tumors (as well as small and even microscopic tumors),without risking damage to non-target tissues, such as skin and normalsubcutaneous tissue. Instead of the conventional technique, a method ofphotoactivation and a series of photosensitizer constructs is neededthat enable PDT induced cytotoxicity, on both a macro and microscopicscale, without risk to the cutaneous layer, or any surrounding normaltissues. Also, the therapeutic index should be enhanced if a specificphotosensitizer drug targeting scheme is employed.

[0023] Citation of the above documents is not intended as an admissionthat any of the foregoing is pertinent prior art. All statements as tothe date or representation as to the contents of these documents isbased on the information available to the applicants and does notconstitute any admission as to the correctness of the dates or contentsof these documents. Further, all documents referred to throughout thisspecification are hereby incorporated by reference herein, in theirentirety.

SUMMARY OF THE INVENTION

[0024] In accord with the present invention, a method is defined fortranscutaneously administering a photodynamic therapy to a target tissuein a mammalian subject. The method includes the step of administering tothe subject a therapeutically effective amount of either aphotosensitizing agent having a characteristic light absorptionwaveband, a photosensitizing agent delivery system that delivers thephotosensitizing agent, or a prodrug that produces a prodrug producthaving a characteristic light absorption waveband. The photosensitizingagent, photosensitizing agent delivery system, or prodrug selectivelybinds to the target tissue. Light having a waveband corresponding atleast in part with the characteristic light absorption waveband of saidphotosensitizing agent or of the prodrug is used for transcutaneouslyirradiating at least a portion of the mammalian subject. An intensity ofthe light used for irradiating is substantially less than 500 mw/cm²,and a total fluence of the light is sufficiently high to activate thephotosensitizing agent or the prodrug product, as applicable.

[0025] Preferably, sufficient time is allowed for any of thephotosensitizing agent, the photosensitizing agent delivery system, orthe prodrug (depending upon which one of these was administered) that isnot bound to the target tissue to clear from non-target tissues of themammalian subject prior to the step of irradiating with the light.

[0026] In one application of the invention, the target tissue isvascular endothelial tissue. In another application, the target tissueis an abnormal vascular wall of a tumor. As further defined, the targettissue is selected from the group consisting of: a vascular endothelialtissue, an abnormal vascular wall of a tumor, a solid tumor, a tumor ofa head, a tumor of a neck, a tumor of a gastrointestinal tract, a tumorof a liver, a tumor of a breast, a tumor of a prostate, a tumor of alung, a nonsolid tumor, malignant cells of one of a hematopoietic tissueand a lymphoid tissue, lesions in a vascular system, a diseased bonemarrow, and diseased cells in which the disease is one of an autoimmuneand an inflammatory disease. In yet a further application of the presentinvention, the target tissue is a lesion in a vascular system. It iscontemplated that the target tissue is a lesion of a type selected fromthe group consisting of atherosclerotic lesions, arteriovenousmalformations, aneurysms, and venous lesions.

[0027] The step of irradiating generally comprises the step of providinga light source that is activated to produce the light. In one preferredembodiment of the invention, the light source is disposed external to anintact skin layer of the mammalian subject during the step ofirradiating. In another preferred embodiment, the method includes thestep of inserting the light source underneath an intact skin layer, butexternal to an intact surface of an organ of the mammalian subject, andthe organ comprises the target tissue.

[0028] Preferably, the photosensitizing agent is conjugated to a ligand.The ligand may be either an antibody or an antibody fragment that isspecific in binding with the target tissue. Alternatively, the ligand isa peptide, or a polymer, either of which is specific in binding with thetarget tissue.

[0029] The photo sensitizing agent is preferably selected from the groupconsisting of indocyanine green (ICG), methylene blue, toluidine blue,aminolevulinic acid (ALA), chlorins, phthalocyanines, porphyrins,purpurins, texaphyrins, and other photosensitizer agents that havecharacteristic light absorption peak in a range of from about 500 nm toabout 1100 nm.

[0030] The step of irradiating is preferably carried out for a timeinterval of from about 30 minutes to about 72 hours, or more preferably,from about 60 minutes to about 48 hours, or most preferably, from about3 hours to about 24 hours.

[0031] In yet another application of the invention, the target tissue isbone marrow, or comprises cells afflicted with either an autoimmunedisease or an inflammatory disease.

[0032] An additional application of the invention contemplates a methodfor administering photodynamic therapy to a target composition in amammalian subject by transillumination. The target composition mayinclude one or more pathogenic agents, including: bacteria, viruses,fungi, protozoa, and toxins as well as tissues infected or infiltratedtherewith.

[0033] Preferably, the total fluence of the light used for irradiatingis between about 30 Joules and about 25,000 Joules, more preferably,between about 100 Joules and about 20,000 Joules, and most preferably,between about 500 Joules and about 10,000 Joules.

[0034] Another application of the present invention uses an energyactivated compound that has a characteristic energy absorption waveband.The energy activated compound selectively binds to the target tissue.Energy having a waveband corresponding at least in part with thecharacteristic energy absorption waveband of said energy activatedcompound is used for transcutaneously irradiating at least a portion ofthe mammalian subject. Preferably the waveband is in the ultrasonicrange of energy. Said compound is activated by said irradiating step,wherein the intensity of said ultrasonic energy is substantially lessthan that level which would result in damage to normal tissue, but at asufficiently high total fluence of ultrasonic energy that is absorbed bysaid compound which in turn destroys the target tissue to which it isbound. Preferably, the total fluence of the ultrasonic energy used forirradiating is between about 5 kHz and more than about 300 MHz, morepreferably, between about 10 kHz and more than about 200 MHz, and mostpreferably, between about 20 kHz and more than about 100 MHz.

BRIEF DESCRIPTION OF THE DRAWING FIGURES

[0035] The foregoing aspects and many of the attendant advantages ofthis invention will become more readily appreciated as the same becomesbetter understood by reference to the following detailed description,when taken in conjunction with the accompanying drawings, wherein:

[0036]FIG. 1 is a schematic diagram illustrating an external lightsource being used to administer transcutaneous cancer-therapy to arelatively large, singular tumor, and to multiple, small tumors;

[0037]FIG. 2 is a schematic cross-sectional view of a section of a tumorblood vessel, illustrating binding of an antibody/photosensitive drug toendothelial tissue;

[0038]FIGS. 3A and 3B are schematic diagrams illustrating biotin-avidintargeting of endothelial antigens for use in rendering PDT;

[0039] FIGS. 4A-4C schematically illustrate tissue amplified infarctiondownstream of photodynamic transcutaneous therapy applied to endotheliumtissue;

[0040]FIG. 5 is a schematic diagram illustrating the use of an externalultrasound source for transcutaneous application of PDT to a deep tumor;

[0041]FIG. 6 is a schematic diagram showing the use of an external lightsource for transcutaneous treatment of intraosseous disease;

[0042]FIG. 7 is a schematic diagram showing both an external lightsource transcutaneously administering light and an intraluminal lightsource position within either the terminal ileum or colon to treatCrohn's disease with targeted PDT;

[0043]FIG. 8 is a schematic diagram illustrating an intraluminal lightsource in the form of a capsule or pill for administering light todestroy H. pylori on the gastric lining with targeted PDT; and

[0044]FIG. 9 is a schematic diagram showing how an internal light sourceadministers transillumination of a deep tumor through an organ wall toprovide targeted PDT that destroys the tumor.

DESCRIPTION OF THE PREFERRED EMBODIMENT Introduction and GeneralDescription of the Invention

[0045] This invention is directed to methods and compositions fortherapeutically treating a target tissue or destroying or impairing atarget cell or a biological component in a mammalian subject by thespecific and selective binding of a photosensitizer agent to the targettissue, cell, or biological component. At least a portion of the subjectis irradiated with light at a wavelength or waveband within acharacteristic absorption waveband of the photosensitizing agent. Thelight is administered at a relatively low fluence rate or intensity, butat an overall high total fluence dose, resulting in minimal collateralnormal tissue damage. It is contemplated that an optimal total fluencefor the light administered to a patient will be determined clinically,using a light dose escalation trial. It is further contemplated that thetotal fluence administered during a treatment will preferably be in therange of 30 Joules to 25,000 Joules, more preferably, in the range from100 Joules to 20,000 Joules, and most preferably, in the range from 500Joules to 10,000 Joules.

[0046] The terminology used herein is generally intended to have the artrecognized meaning and any differences therefrom as used in the presentdisclosure, will be apparent to the ordinary skilled artisan. For thesake of clarity, terms may also have a particular meaning, as will beclear from their use in context. For example, “transcutaneous” as usedin regard to light in this specification and in the claims that follow,more specifically herein refers to the passage of light through unbrokentissue. Where the tissue layer is skin or dermis, transcutaneousincludes “transdermal” and it will be understood that the light sourceis external to the outer skin layer. However, the term“transillumination” as used herein refers to the passage of lightthrough a tissue layer, such as the outer surface layer of an organ,e.g., the liver, and it will be apparent that the light source isexternal to the organ, but internal or implanted within the subject orpatient.

[0047] One aspect of the present invention provides for the precisetargeting of photosensitive agents or drugs and compounds to specifictarget antigens of a subject or patient and to the method for activatingthe targeted photosensitizer agents by subsequently administering to thesubject light at a relatively low fluence rate or intensity, over aprolonged period of time, from a light source that is external to thetarget tissue in order to achieve maximal cytotoxicity of the abnormaltissue, with minimal adverse side effects or collateral normal tissuedamage.

[0048]FIG. 1 illustrates transcutaneous delivery of light 12 from anexternal source 10 to a relatively deep tumor 14, or to a plurality ofsmall, but relatively deep tumors 16. The light emitted by externalsource 10 is preferably of a longer waveband, but still within anabsorption waveband of the photosensitive agent (not shown in thisFigure) that has been selectively bound to tumor 14 and smaller tumors16. The longer wavelength of light 12 enables it to pass through adermal layer 18 and penetrate into the patient's body beyond the depthof tumor(s) being treated with targeted PDT. In these two examples, thePDT is directed specifically at target cells in tumor 14 or in tumors16.

[0049] As used in this specification and the following claims, the terms“target cells” or “target tissues” refer to those cells or tissues,respectively that are intended to be impaired or destroyed by PDTdelivered in accord with the present invention. Target cells or targettissues take up or bind with the photosensitizing agent, and, whensufficient light radiation of the waveband corresponding to thecharacteristic waveband of the photosensitizing agent is applied, thesecells or tissues are impaired or destroyed. Target cells are cells intarget tissue, and the target tissue includes, but is not limited to,vascular endothelial tissue, abnormal vascular walls of tumors, solidtumors such as (but not limited to) tumors of the head and neck, tumorsof the gastrointestinal tract, tumors of the liver, tumors of thebreast, tumors of the prostate, tumors of the lung, nonsolid tumors andmalignant cells of the hematopoietic and lymphoid tissue, other lesionsin the vascular system, bone marrow, and tissue or cells related toautoimmune disease.

[0050] Further, target cells include virus-containing cells, andparasite-containing cells. Also included among target cells are cellsundergoing substantially more rapid division as compared to non-targetcells. The term “target cells” also includes, but is not limited to,microorganisms such as bacteria, viruses, fungi, parasites, andinfectious agents. Thus, the term “target cell” is not limited to livingcells but also includes infectious organic particles such as viruses.“Target compositions” or “target biological components” include, but arenot be limited to: toxins, peptides, polymers, and other compounds thatmay be selectively and specifically identified as an organic target thatis intended to be impaired or destroyed by this treatment method.

[0051]FIG. 2 includes a section of a tumor blood vessel 20 having a wall22, with an endothelial lining 24. A plurality of endothelial antigens26 are disposed along the endothelial lining. In this example,antibodies 28 that are specific to endothelial antigens 26 have beenadministered and are shown binding with the endothelial antigens.Coupled to antibodies 28 are PDT photosensitive drug molecules 30. Thus,the PDT photosensitive drug molecules are bound to the endothelialantigens via antibodies 28, but are not bound to non-target cells, sincethe antibodies are selective only to the endothelial antigens.

[0052] “Non-target cells” are all the cells of a mammal that are notintended to be impaired, damaged, or destroyed by the treatment methodrendered in accord with the present invention. These non-target cellsinclude but are not limited to healthy blood cells, and other normaltissue, not otherwise identified to be targeted.

[0053] “Destroy” means to kill the desired target cell. “Impair” meansto change the target cell in such a way as to interfere with itsfunction. For example, in North et al., it is observed that aftervirus-infected T cells treated with benzoporphyrin derivatives (“BPD”)were exposed to light, holes developed in the T cell membrane andincreased in size until the membrane completely decomposed (Blood Cells18:129-40, (1992)). Target cells are understood to be impaired ordestroyed even if the target cells are ultimately disposed of bymacrophages.

[0054] “Energy activated agent” is a chemical compound that binds to oneor more types of selected target cells and, when exposed to energy of anappropriate waveband, absorbs the energy, causing substances to beproduced that impair or destroy the target cells.

[0055] “Photosensitizing agent” is a chemical compound that binds to oneor more types of selected target cells and, when exposed to light of anappropriate waveband, absorbs the light, causing substances to beproduced that impair or destroy the target cells. Virtually any chemicalcompound that preferentially is absorbed or bound to a selected targetand absorbs light causing the desired therapy to be effected may be usedin this invention. Preferably, the photosensitizing agent or compound isnontoxic to the animal to which it is administered or is capable ofbeing formulated in a nontoxic composition that can be administered tothe animal. In addition, following exposure to light, thephotosensitizing agent in any resulting photodegraded form is alsopreferably nontoxic. A comprehensive listing of photosensitive chemicalsmay be found in Kreimer-Bimbaum, Sem. Hematol, 26:157-73, (1989).Photosensitive agents or compounds include, but are not limited to,chlorins, bacteriochlorins, phthalocyanines, porphyrins, purpurins,merocyanines, psoralens, benzoporphyrin derivatives (BPD), and porfimersodium and pro-drugs such as delta-aminolevulinic acid, which canproduce photosensitive agents such as protoporphyrin IX. Other suitablephotosensitive compounds include ICG, methylene blue, toluidine blue,texaphyrins, and any other agent that absorbs light in a range of 500nm-1100 nm.

[0056] The term “prodrug” is used herein to mean any of a class ofsubstances that are not themselves photosensitive agents, but whenintroduced into the body, through metabolic, chemical, or physicalprocesses, are converted into a photosensitive agent. In the followingdisclosure, an aminolevulinic acid (ALA) is the only exemplary prodrug.After being administered to a patient, ALA is metabolically convertedinto a porphyrin compound that is an effective photosensitive agent.

[0057] “Radiation” as used herein includes all wavelengths andwavebands. Preferably, the radiation wavelength or waveband is selectedto correspond with or at least overlap the wavelength(s) or wavebandsthat excite the photosensitive compound. Photosensitive agents orcompound typically have one or more absorption wavebands that excitethem to produce the substances, which damage or destroy target tissue,target cells, or target compositions. Even more preferably, theradiation wavelength or waveband matches the excitation wavelength orwaveband of the photosensitive compound and has low absorption by thenon-target cells and the rest of the intact animal, including bloodproteins. For example, a preferred wave length of light for ICG is inthe range 750-850 nm.

[0058] The radiation used to activate the photosensitive compound isfurther defined in this invention by its intensity, duration, and timingwith respect to dosing a target site. The intensity or fluence rate mustbe sufficient for the radiation to penetrate skin and reach the targetcells, target tissues, or target compositions. The duration or totalfluence dose must be sufficient to photoactivate enough photosensitiveagent to achieve the desired effect on the target site. Both intensityand duration are preferably limited to avoid over treating the subjector animal. Timing with respect to the dosage of the photosensitive agentemployed is important, because (1) the administered photosensitive agentrequires some time to home in on target cells, tissue, or compositionsat the treatment site, and (2) the blood level of many photosensitiveagents decreases with time.

[0059] The present invention provides a method for providing a medicaltherapy to an animal, and the term “animal” includes, but is not limitedto, humans and other mammals. The term “mammals” or “mammalian subject”includes farm animals, such as cows, hogs and sheep, as well as pet orsport animals such as horses, dogs, and cats.

[0060] Reference herein to “intact animal” means that the whole,undivided animal is available to be exposed to radiation. No part of theanimal is removed for exposure to the radiation, in contrast withphotophoresis, in which an animal's blood is circulated outside its bodyfor exposure to radiation. However, in the present invention, the entireanimal need not be exposed to radiation. Only a portion of the intactanimal subject may or need be exposed to radiation, sufficient to ensurethat the radiation is administered to the treatment site where thetarget tissue, cells, or compositions are disposed.

[0061] In the present invention, a photosensitizing agent is generallyadministered to the animal before the animal is subjected to radiation.Preferred photosensitizing agents include, but are not limited to,chlorins, bacteriochlorins, phthalocyanines, porphyrins, purpurins,merocyanines, psoralens and pro-drugs such as delta.-aminolevulinicacid, which can produce drugs such as protoporphyrin. More preferredphotosensitizing agents are: methylene blue, toluidine blue,texaphyrins, and any other agent that absorbs light having a wavelengthor waveband in the range from 600 nm-1100 nm. Most preferred of thephotosensitizing agents is ICG. The photosensitizing agent is preferablyadministered locally or systemically, by oral ingestion, or byinjection, which may be intravascular, subcutaneous, intramuscular,intraperitoneal or directly into a treatment site, such as intratumoral.The photosensitizing agent also can be administered enterally ortopically via patches or implants.

[0062] The photosensitizing agent also can be conjugated to specificligands known to be reactive with a target tissue, cell, or composition,such as receptor-specific ligands or immunoglobulins or immunospecificportions of immunoglobulins, permitting them to be more concentrated ina desired target cell or microorganism than in non-target tissue orcells. The photosensitizing agent may be further conjugated to aligand-receptor binding pair. Examples of a suitable binding pairinclude but are not limited to: biotin-streptavidin, chemokine-chemokinereceptor, growth factor-growth factor receptor, and antigen-antibody. Asused herein, the term “photosensitizing agent delivery system” refers toa photosensitizing agent conjugate, which because of its conjugation,has increased selectivity in binding to a target tissue, target cells,or target composition. The use of a photosensitizing agent deliverysystem is expected to reduce the required dose level of the conjugatedphotosensitizing agent, since the conjugate material is more selectivelytargeted at the desired tissue, cell, or composition, and less of it iswasted by distribution into other tissues whose destruction should beavoided.

[0063] In FIGS. 3A and 3B, an example of a photosensitizing agentdelivery system 40 is illustrated in which the target tissue isendothelial layer 24, which is disposed along blood vessel wall 22 oftumor blood vessel 20. As shown in FIG. 3A, antibodies 28 are coupledwith biotin molecules 42 and thus selectively bound to endothelialantigens 26 along the endothelial layer. FIG. 3B illustrates avidinmolecules 44 coupled to PDT photosensitive drug molecules 30, where theavidin molecules bind with biotin molecules 42. This system thus ensuresthat the PDT photosensitive drug molecules 30 only bind with theselectively targeted endothelial tissue. When light of the appropriatewaveband is administered, it activates the PDT photosensitive drugmolecules, causing the endothelial tissue to be destroyed.

[0064] FIGS. 4A-4C illustrate a mechanism for amplifying the effect on atumor of PDT administered to destroy the endothelial tissue in a tumorblood vessel 50. Tumor blood vessel 50 distally branches into twosmaller blood vessels 52. In FIG. 4A, the PDT administered to activatethe PDT photosensitive drug molecules has produced substantial damage tothe endothelium, creating an intravascular thrombosis (or clot) 54. Asshown in FIG. 4B, the intravascular thrombosis is carried distallythrough tumor blood vessel 50 until it reaches the bifurcation pointwhere smaller diameter blood vessels 52 branch. Due to the flow throughsmaller internal diameter of blood vessels 52, intravascular thrombosis54 can not advance any further, and is stopped, creating a plug thatvirtually stops blood flow through tumor blood vessel 50. Theinterruption of blood flow also interrupts the provision of nutrientsand oxygen to the surrounding tumor cells, causing the tumor cells todie. In FIG. 4C, the dying tumor cells 56 are within a zone of tumorcell death or necrosis 58 surrounding the vessel and which zoneincreases in volume over time, thereby amplifying the effects of the PDTon the endothelium tissue of the tumor blood vessels.

[0065] A photosensitizing agent can be administered in a dryformulation, such as pills, capsules, suppositories or patches. Thephotosensitizing agent also may be administered in a liquid formulation,either alone, with water, or with pharmaceutically acceptableexcipients, such as are disclosed in Remington's PharmaceuticalSciences. The liquid formulation also can be a suspension or anemulsion. In particular, liposomal or lipophilic formulations aredesirable. If suspensions or emulsions are utilized, suitable excipientsinclude water, saline, dextrose, glycerol, and the like. Thesecompositions may contain minor amounts of nontoxic auxiliary substancessuch as wetting or emulsifying agents, antioxidants, pH bufferingagents, and the like.

[0066] The dose of photosensitizing agent will vary with the targettissue, cells, or composition, the optimal blood level (see Example 1),the animal's weight, and the timing and duration of the radiationadministered. Depending on the photosensitizing agent used, anequivalent optimal therapeutic level will have to be empiricallyestablished. Preferably, the dose will be calculated to obtain a desiredblood level of the photosensitizing agent, which will likely be betweenabout 0.01 μg/ml and 100 μg/ml. More preferably, the dose will produce ablood level of the photosensitizing agent between about 0.01 μg/ml and10 μg/ml.

[0067] The intensity of radiation used to treat the target cell ortarget tissue is preferably between about 5 mW/cm² and about 100 mW/cm².More preferably, the intensity of radiation employed should be betweenabout 10 mW/cm² and about 75 mW/cm². Most preferably, the intensity ofradiation is between about 15 mW/cm² and about 50 mW/cm².

[0068] The duration of radiation exposure administered to a subject ispreferably between about 30 minutes and about 72 hours. More preferably,the duration of radiation exposure is between about 60 minutes and about48 hours. Most preferably, the duration of radiation exposure is betweenabout 2 hours and about 24 hours.

[0069] It is contemplated that a targeted photosensitizer agent can besubstantially and selectively photoactivated in the target cells andtarget tissues within a therapeutically reasonable period of time andwithout excess toxicity or collateral damage to non-target normaltissues. Thus, there appears to be a therapeutic window bounded by thetargeted photosensitizer agent dosage and the radiation dosage. In viewof problems in the prior art related to either extracorporeal treatmentof target tissues or use of high intensity laser light irradiationintra-operatively, the present invention offers substantial advantages.In accord with the present invention, targeted transcutaneous PDT willbe employed to treat patients injected with a photosensitizer agent andwill subject the patients to a relatively low fluence rate, but hightotal fluence dose of radiation. This approach is an attractive methodfor treating target tissues that include neoplastic diseased tissue,infectious agents, and other pathological tissues, cells, andcompositions.

[0070] One aspect of the present invention is drawn to a method fortranscutaneous energy activation therapy applied to destroy tumors in amammalian subject or patient by first administering to the subject atherapeutically effective amount of a first conjugate comprising a firstmember of a ligand-receptor binding pair conjugated to an antibody orantibody fragment. The antibody or antibody fragment selectively bindsto a target tissue antigen. Simultaneously or subsequently, atherapeutically effective amount of a second conjugate comprising asecond member of the ligand-receptor binding pair conjugated to anenergy-sensitive agent or energy-sensitive agent delivery system orprodrug is administered to the patient, wherein the first member bindsto the second member of the ligand-receptor binding pair. These stepsare followed by irradiating at least a portion of the subject withenergy having a wavelength or waveband absorbed by the energy-sensitiveagent, or energy-sensitive agent delivery system, or by the productthereof. This radiation energy is preferably provided by an energysource that is external to the subject and is preferably administered ata relatively low fluence rate that results in the activation of theenergy-sensitive agent, or energy-sensitive delivery system, or prodrugproduct.

[0071] While one preferred embodiment of the present invention is drawnto the use of light energy for administering PDT to destroy tumors,other forms of energy are within the scope of this invention, as will beunderstood by those of ordinary skill in the art. Such forms of energyinclude, but are not limited to: thermal, sonic, ultrasonic, chemical,light, microwave, ionizing (such as x-ray and gamma ray), mechanical,and electrical. For example, sonodynamically induced or activated agentsinclude, but are not limited to: gallium-porphyrin complex (see Yumitaet al., Cancer Letters, 112: 79-86, (1997)), other porphyrin complexes,such as protoporphyrin and hematoporphyrin (see Umemura et al.,Ultrasonics Sonochemistry 3:S187-S191, (1996)); other cancer drugs, suchas daunorubicin and adriamycin, used in the presence of ultrasoundtherapy (see Yumita et al., Japan J. Hyperthermic Oncology,3(2):175-182, (1987)).

[0072]FIG. 5 illustrates the use of an external ultrasound transducerhead 60 for generating an ultrasonic beam 62 that penetrates through adermal layer 64 and into a subcutaneous layer 66. The externalultrasound transducer head is brought into contact with dermal layer 64so that ultrasonic beam 62 is directed toward a relatively deep tumor68. The ultrasonic beam activates a PDT photosensitive drug that hasbeen administered to the patient and selectively targeted at tumor 68,causing the drug to destroy the tumor.

[0073] This invention further preferably employs an energy source, e.g.,a light source, that is external to the target tissue. The targettissues may include and may relate to the vasculature or blood vesselsthat supply blood to tumor tissue or the target tissues may include thetumor tissue antigens, per se. These target tissue antigens will bereadily understood by one of ordinary skill in the art to include but tonot be limited to: tumor surface antigen, tumor endothelial antigen,non-tumor endothelial antigen, and tumor vessel wall antigen, or otherantigens of blood vessels that supply blood to the tumor.

[0074] Where the target tissue includes endothelial or vascular tissue,a preferable ligand-receptor binding pair includes biotin-streptavidin.In this preferred embodiment, the activation of photosensitizer agentsby a relatively low fluence rate of a light source over a prolongedperiod of time results in the direct or indirect destruction, impairmentor occlusion of blood supply to the tumor resulting in hypoxia or anoxiato the tumor tissues. Where the target tissue includes tumor tissueother than endothelial or vascular, the activation of photosensitizeragents by a relatively low fluence rate of a light source over aprolonged period of time results in the direct destruction of the tumortissue due to deprivation of oxygen and nutrients from the tumor cells.

[0075] The ordinary skilled artisan would be familiar with variousligand-receptor binding pairs, including those known and those currentlyyet to be discovered. Those known include, but are not limited to:biotin-streptavidin, chemokine-chemokine receptor, growth factor-growthfactor receptor, and antigen-antibody. The present inventioncontemplates at least one preferred embodiment that usesbiotin-streptavidin as, the ligand-receptor binding pair. However, theordinary skilled artisan will readily understand from the presentdisclosure that any ligand-receptor binding pair may be useful inpracticing this invention, provided that the ligand-receptor bindingpair demonstrates a specificity for the binding by the ligand to thereceptor and further provided that the ligand-receptor binding pairpermits the creation of a first conjugate comprising a first member ofthe ligand-receptor binding pair conjugated to an antibody or antibodyfragment. In this case, the antibody or antibody fragment selectivelybinds to a target tissue antigen and permits the creation of a secondconjugate comprising a second member of the ligand-receptor binding pairconjugated to an energy-sensitive or photosensitizing agent, orenergy-sensitive or photosensitizing agent delivery system, or prodrug.The first member then binds to the second member of the ligand-receptorbinding pair.

[0076] Another preferred embodiment of the present invention includes aphotosensitizing agent delivery system that utilizes both a liposomedelivery system and a photosensitizing agent, where each is separatelyconjugated to a second member of the ligand-receptor binding pair, andwhere the first member binds to the second member of the ligand-receptorbinding pair. More preferably, the ligand-receptor binding pair isbiotin-streptavidin. In this embodiment, the photosensitizing agent aswell as the photosensitizing agent delivery system may both bespecifically targeted through selective binding to a target tissueantigen by the antibody or antibody fragment of the first member bindingpair. Such dual targeting is expected to enhance the specificity ofuptake and to increase the quantity of uptake of the photosensitizingagent by the target tissue, cell, or compositions.

EXAMPLES

[0077] Having now generally described the invention, it will be morereadily understood through reference to the following examples, whichare provided by way of illustration and are not intended to be limitingin regard to the scope of the invention, unless specified.

Example 1

[0078] Transcutaneous Photodynamic Therapy of a Solid Type Tumor

[0079] A patient in the terminal phase of recurrent malignant coloncancer presented with a protruding colon carcinoma tumor mass ofapproximately 500 grams and approximately 13 cm in diameter, whichextended through the patient's dermis. Due to the advanced state of thepatient's disease and due to the highly vascularized nature of thistumor mass, resection was not feasible. Further, this large tumor masspresented a significant amount of pain and discomfort to the patient, aswell as greatly impairing the patient's ability to lie flat.

[0080] Six separate light source probes, each including a linear arrayof LEDs, were surgically implanted in this large human tumor usingstandard surgical procedures. An intensity of about 25-30 mW of lightfrom each light source probe (650 nm peak wavelength) was delivered tothe tumor for 40 hours following oral administration to the patient of asingle dose of an (ALA) photosensitizer agent (60 mg/kg). However, after18 hours, two of the light source probes became unseated from the tumormass and were disconnected from the electrical power supply used toenergize the LEDs on each probe. The total fluence delivered to thetumor bed during this single extended duration treatment was in excessof 20,000 Joules. Extensive tumor necrosis in a radius of up to 5 cmaround each of the light source probes was observed after 40 hours ofPDT, with no collateral damage to surrounding normal tissue. The extentof this PDT induced necrotic effect in a large volume of tumor tissuewas totally unexpected and has not been described before in any PDTstudies in subjects in vivo or clinically. Over the course of four weeksfollowing PDT, the necrotic tumor tissue was debrided from the patientresulting in a reduction of approximately 500 grams of tumor tissue. Thepatient noted a significant improvement in his quality of life, with aresurgent level of energy and improved well being.

[0081] The average thickness of human skin is approximately 1 cm.Therefore, if this same method of prolonged, relatively low fluencerate, but overall high total fluence of light delivery is utilized todeliver the light transcutaneously, a therapeutic effect well below theskin surface, to a depth of is contemplated.

[0082] The fluence rate employed in this Example represented about150-180 mW/cm², with a total fluence more than 20,000 Joules. Thepreferable fluence rate contemplated more broadly by the presentinvention is between about 5 mW/cm² and about 100 mW/cm², morepreferably, between about 10 mW/cm² and about 75 mW/cm², and mostpreferably, between about 15 mW/cm² and about 50 mW/cm².

[0083] It is further contemplated that the optimal total fluence beempirically determined, using,a light dose escalation trial, and willlikely and preferably be in the range of about 30 Joules to about 25,000Joules, and more preferably be in the range from about 100 Joules toabout 20,000 Joules, and most preferably be in the range from about 500Joules to about 10,000 Joules.

Example 2

[0084] Transcutaneous Photodynamic Therapy of Intraosseous Disease

[0085] The current accepted therapy for treating leukemia and othermalignant bone marrow diseases employs a systemic treatment utilizingchemotherapy and/or radiotherapy, sometimes followed by a bone marrowtransplant. There are significant risks associated withnon-discriminative ablative therapies that destroy all marrow elements,including the risks of infections, bleeding diathesis, and otherhematological problems.

[0086] There is a definite need for alternative therapies that do notsubject patients to procedures which may be risky and which inherentlycause pain and suffering. This example is directed to a method oftreating intraosseous malignancy that has major advantages over theprior art techniques for treating this disease.

[0087] A targeted antibody-photosensitizer conjugate (APC) isconstructed, which binds selectively to antigens present on leukemiccells. This ligand-receptor binding pair or APC is infused intravenouslyand is taken up in the marrow by circulating leukemic cells, and bystationary deposits that may reside in other organs. When unbound toleukemic cells, APC is eliminated from the body. Internal or externallight sources may be used to activate the targeted drug. For example,light bar probes disclosed in U.S. Pat. No. 5,445,608 may be insertedinto bone marrow to treat the intraosseous disease. The devicesdisclosed in U.S. Pat. No. 5,702,432 may be used to treat disease cellscirculating in the patient's lymphatic or vascular system. An externaldevice transcutaneously activating the targeted drug, for example, alight source that emits light that is transmitted through the dermallayer may also be used in treating the marrow compartment in accord withthe present invention.

[0088] PDT targeting has been described for leukemic cells (see U.S.Pat. No. 5,736,563). but not with capability of treating marrow in situ.Without this capability, simply lowering the leukemic cell count wouldhave little clinical benefit, since the marrow is a major source of newleukemic clones, and the marrow must. be protected from failure, whichwill lead to the death of the patient regardless of how well thepathologic cell load in the circulation is treated. Specific APCpromotes the selective damage of leukemic cells in marrow, whilereducing collateral and non-target tissue damage. Further, the use of arelatively low fluence rate, but overall high total fluence dose isparticularly effective in this therapy. Optimal fluence rates and dosingtimes are readily empirically determined using dose escalation for bothdrug and light dose as is often done in a clinical trial. Any of anumber of different types of leukemia cell antigens may be selected,provided that the antigen chosen is as specific as possible for theleukemia cell. Such antigens will be known to those of ordinary skill inthis art. The selection of a specific photosensitizer agent may be made,provided that the photosensitizer agent chosen is activated by lighthaving a waveband of from about 500 nm to about 1100 nm, and morepreferably, a waveband from about 630 nm to about 1000 nm, and mostpreferably, a waveband from about 800 nm to about 950 nm or greater. Thephotosensitizer agents noted above are suitable for use in this Example.

[0089] With reference to FIG. 6, external light source 10 isadministering light 12 transcutaneously through dermal layer 18. Light12 has a sufficiently long wavelength to pass through a subcutaneouslayer 70 and through a cortical bone surface 74, into a bone marrowcompartment 76. Leukemia cells 78 have penetrated bone marrowcompartment 76 and are distributed about within it. To provide targetedPDT treatment that will destroy the leukemia cells, antibodies 82 boundwith PDT photosensitive drug molecules 84 have been administered to thepatient and have coupled with leukemia antigens 80 on the leukemia cells78. The light provided by external light source 10 thus activates thePDT photosensitive drug, causing it to destroy the leukemia cells. Thistargeted PDT process is carried out with minimal invasive or adverseimpact on the patient, in contrast to the more conventional treatmentparadigms currently used.

Example 3

[0090] Transcutaneous Photodynamic Therapy of Crohn's Disease

[0091] Crohn's disease is a chronic inflammation of the gastrointestinaltract thought to be mediated in large part by dysfunction of CD4⁺ Tcells lining the gut mucosa, especially in terminal ileum. The currentaccepted therapy for Crohn's disease provides for surgical removal ofthe inflamed bowel segment and the use of anti-inflammatory agents,steroids and other immunosuppressive drugs. None of these measures isentirely satisfactory due to surgical risk, recurrence of disease,medication side effects, and refractoriness of the disease. There is aclear need for alternative therapies useful in treating this immunedysfunction that offer greater efficacy and reduced side effects andrisk. This Example, details of which are illustrated in FIG. 7,indicates the drug compositions and methodologies useful in accord withthe present invention to selectively destroy the dysfunctional cells orinhibit their function. In the illustrated example, external lightsource 10 is administering light 12 that has a sufficiently longwavelength to penetrate dermal tissue 18, which is disposed over apatient's abdomen, and pass through a subcutaneous layer 90, into aterminal ileum or colon 92. The light passes through wall 94 of theterminal ileum or colon. Alternatively (or in addition), light 12′ canbe administered from an intraluminal probe 96, from sources (notseparately shown) that are energized with an electrical current suppliedthrough a lead 98.

[0092] Ligand-receptor binding pairs 100, or more specifically, APCs,are created that bind selectively to CD4⁺ T cell antigens 102 of T cells104, which are disposed along the interior, intraluminal surface of theterminal ileum or colon. For example, the CD4⁺ antigen itself may betargeted by those antibodies 106 that bind specifically to the CD4⁺antigen. Many of the photosensitizer agents noted above may be used forphotosensitizing drug molecules 108, in the therapy of this Example. TheAPC is preferably formulated into a pharmaceutically acceptable compoundthat can be released in the terminal ileum and colon in a manner similarto that known to be used for the orally delivered form of Budesonide™also known as Entocort™. The APC, compound is ingested and releases theconjugate into the terminal ileum and colon. At the time of therapy, thebowel should have been prepped in much the same manner as done inpreparing for a colonoscopy, so that it is cleared of fecal material.The targeted photosensitizer will bind to the pathologic T cells and anyunbound APC is removed via peristaltic action. The sensitizer bound tothe T cells is activated by intraluminally positioned light source probe96, details of which are disclosed in any one of U.S. Pat. Nos.5,766,234; 5,782,896; 5,800,478; and 5,827,186, each of which is herebyincorporated by reference herein in its entirety; or by a flexibleintraluminal optical fiber (not shown) that is passed via thenasopharynx; or, by the transcutaneous light illumination provided byexternal light source 10. Transcutaneous light illumination is preferredbecause it is entirely noninvasive.

[0093] In this exemplary treatment, the following protocol may beutilized:

[0094] Step 1 Patient is NPO (“non per os” or nothing by mouth) and thebowel has been prepped or cleansed by administering an enema to clear itof fecal material;

[0095] Step 2 Specially formulated APC conjugate compound 100 isingested;

[0096] Step 3 The APC conjugate is released to the terminal ileum andcolon;

[0097] Step 4 If transcutaneous illumination is not used, one or morelight source probes 96 are ingested or passed into the GI tract andadvanced to the terminal ileum or colon.

[0098] Step 5 the APC conjugate is bound to target T cells 104 and anyunbound conjugate fraction passes distally via peristalsis (and issubsequently eliminated from the body).

[0099] Step 6 If an internal light source is used, the light sourceshould preferably be imaged using ultrasound or computer assistedtopography (i.e., a CT scan—not shown) to confirm—its location and the Ilight source can then be activated while positioned in the ileum. Onceactivated, the light source will deliver light at the appropriatewaveband for the photosensitizing agent selected, at a relatively lowfluence rate, but at a high total fluence dose, as noted above. Theoptimal drug dose and fluence parameters will be determined clinicallyin a drug and light dose escalation trial. The light dose and drug doseare such that T cell inactivation occurs, leading to decreasedregulation of the immune process and a reduction of any pathologicinflammation—both of which are factors—characteristic of this disease.

[0100] Step 7 The light source is deactivated. It is particularlyimportant to deactivate an internal light source before withdrawing itfrom the treatment site to prevent nonspecific APC activation.

[0101] The present invention can also be employed to target other typesof immunologic cells, such as other T cells, macrophages, neutrophils, Bcells, and monocytes. A tiered approach can thus be employed, startingwith CD4⁺ T cells, then moving to CD8⁺ T cells, and then monocytes, andneutrophils. By inhibiting or preventing interaction and/or secretion ofinflammatory cell products, the pathologic process is controlled at thelumenal site, completely avoiding systemic side effects and majorsurgery. The same process can be applied to treat ulcerative colitiswith the same benefits. As indicated above, the APC can be activatedwith light administered transcutaneously, using any number of differenttypes of external light sources such as LEDs, laser diodes, and lampsthat emit light with a wavelength or waveband sufficiently long topenetrate through the overlying dermal and internal tissue, and into theintestine. The, optimal wavelength or waveband of this light isdetermined by both the light absorption properties of thephotosensitizer and the need to use light with as long a wavelength aspossible to ensure adequate penetration into the patient's body. Adesirable photosensitizer is preferably one that absorbs in the rangefrom about 700 nm to about 900 rim, which optimizes tissue penetration.The appropriate fluence rate and total fluence delivered is readilydetermined by a light dose escalation clinical trial. The light dose anddrug dose are such that T cell inactivation occurs, leading to reducedregulation of the immune process and a reduction in pathologicinflammation.

Example 4

[0102] Intraluminal Transcutaneous PDT Targeted at Helicobacterpylori

[0103] Targeting of photosensitizers to bind with bacterial cells isknown in the prior art. Many antigens that can serve as targets forligand-receptor binding pairs, and more specifically, APC have beenidentified, and the techniques to construct such conjugates are wellknown to those of ordinary skill in this art. What is not apparent fromthe prior art are the steps necessary to for apply such conjugates inthe treatment of a clinical disease. This Example describes the clinicalapplication of APC to the treatment of an infection using PDT. FIG. 8illustrates details of the example, as described below.

[0104]Helicobacter pylori is reportedly associated with tumors of thestomach in mice and as a putative agent of ulcerative pathology inhumans. However, it appears that the use of PDT for destroying an H.pylori infection in human patients has not been carried out, althoughproposals to use laser light for PDT destruction of bacteria have beenset forth (Millson et al., J. of Photochemistry and Photobiology, 32:59-65 (1996)).

[0105] In this Example, a capsular or pill-shaped and sized light source120 is administered orally to a patient, so that it passes into thestomach 118 of the patient, where it administers light 122.Alternatively, an optical fiber (not shown) may be passed into thestomach via the nasopharynx to administer light 122 to the treatmentsite. In order to implement targeted PDT for treating ulcers in humans,an APC 124, which antibody 131 is targeted against a suitableHelicobacter pylori antigen 126 is formulated into an ingestiblecompound that releases the APC to a gastric mucus/epithelial layer 128where the bacterium is found. The APC is ingested at a time when thestomach and duodenum is substantially empty in order to promote bindingof the APC to bacterium 130. Any unbound APC is diluted by gastric juiceand carried distally by peristalsis to be eliminated from the body infecal matter. Light sources suitable for intraluminal passage aredisclosed in any one of U.S. Pat. Nos. 5,766,234; 5,782,896; 5,800,478;and 5,827,186, the disclosure of each being specifically herebyincorporated herein in its entirety. Alternatively, light source 120 incapsule or pill form, e.g., as disclosed in copending commonly assignedU.S. patent application, Ser. No. 09/260,923, entitled, “Polymer Batteryfor Internal Light Device.”—filed on Mar. 2, 1999 is used for activatingthe APC. The light source is preferably energized just prior to itsingestion or remotely after ingestion, when in the stomach or in adesired intraluminal passage. If necessary, multiple light sources areingested to insure that adequate photoactivation of the localized APCoccurs sufficient to kill the bacterium. Light is delivered at arelatively low fluence rate but at a high total fluence dose, asdiscussed above. The light source(s) may be deactivated after passagebeyond the duodenum to avoid unwanted distal photoactivation. In thismanner, a photosensitizing agent 132 comprising the APC is activatedtopically without the need for a procedure such as endoscopy withfiberoptic gastric illumination in order to provide the activatinglight. Since the APC is targeted, nonspecific uptake by normal tissueand other normal compositions of the body is minimized in order toprevent injury to normal gastric tissue and problems with the gastricsystem.

[0106] In this exemplary treatment, the following protocol may beutilized:

[0107] Step 1 Patient is NPO for six hours to insure that the stomach isempty.

[0108] Step 2 The APC is ingested.

[0109] Step 3 One hour elapses to allow for bacterial binding and distalpassage of unbound APC. The optimal period can be longer or shorter andis readily determined by measuring the clinical response; for example,response can be determined endoscopically by observation and biopsy.

[0110] Step 4 One or more light sources are ingested sequentially andactivated in the stomach. The length of time that light is administeredby these sources and the number of sources that are ingested will bedetermined clinically in a light dose escalation study. The churningaction of the stomach serves to translocate the light source(s) so thatthe light is distributed more evenly prior to passage of the source(s)into the duodenum. Since each light source is small (the size of a pillor tablet), it passes easily out through the GI system via peristalsis.

[0111] Step 5 The light sources are deactivated after distal passagebeyond the gastroduodenal area and excreted in fecal matter.

[0112] Note that it is also contemplated that an external light sourcelocated over the gastric area can be used to transcutaneously administerlight to the treatment site, and that an ultrasonic transducer (notshown here, but generally like that shown in FIG. 5) can alternativelybe employed to activate the APC, provided that photosensitizer agent 132comprising the APC is activated by the frequency of ultrasonic energytransmitted by the transducer. The use of an external light sourcerequires that the APC and the light source absorb and emit in the nearinfrared to infrared range, respectively, so that the light willefficiently penetrate the patient's skin and reach the treatment site.Examples of long waveband photosensitizers are ICG, toluidine blue, andmethylene blue, as disclosed herein.

Example 5

[0113] Transcutaneous PDT for Targeting Pulmonary Tuberculosis

[0114] An APC is formulated to bind with great affinity to Mycobacteriumtuberculosis in a selective and specific manner. Preferably, the APC isformulated as an aerosol, which can be easily inhaled, enablingdistribution into all lung segments. Steam is then inhaled to solubilizeany unbound APC and facilitate its removal from the lung by exhalation.Alternatively, the APC is formulated as an injectable compound andadministered intravenously. Either way, the bound APC is photoactivatedby an external light source disposed on the chest and/or back.

[0115] Step 1 The APC is inhaled or injected.

[0116] Step 2 Time is allowed to elapse to allow binding of the APC withthe Mycobacterium tuberculosis, followed by steam inhalation to removeany unbound APC (if inhaled). The time required to ensure atherapeutically effective dose of bound APC may be routinely determinedclinically using standard clinical practices and procedures.

[0117] Step 3 The light source is disposed adjacent to the thorax andactivated for a sufficient time to ensure that therapeutic irradiationhas occurred, which may be routinely determined clinically usingconventional clinical practices and procedures. The fluence rate andtotal fluence dose may be determined as noted above.

[0118] Note that alternatively, an internal light source disposed withinthe thoracic area can be used to administer the light. A furtheralternative would be the use of an external ultrasonic transducer toproduce ultrasonic sound waves that activate the APC. The use of anexternal light source requires that the APC and the light sourcerespectively absorb and emit light in the near infrared to infraredrange to ensure efficient skin penetration of the light. Examples oflong waveband photosensitizers are ICG, toluidine blue, methylene blue.

Example 6

[0119] Transcutaneous PDT for Targeting Otitis Media

[0120] A photosensitizer conjugate is formulated which binds with greataffinity to Streptococcus pneumoniae and Hemophilus influenzae in aselective manner. The APC is formulated into an injectable compound,which can be administered intravenously or instilled topically into themiddle ear via a previously placed tympanostomy tube. The drug isactivated using light emitted by a small light source about the size,shape, and weight of a hearing aid, which is disposed behind the ear andaimed at the middle ear, so that the light passes into the middle eartranscutaneously.

[0121] Step 1 The APC fluid formulation is instilled into the middleear.

[0122] Step 2 Sufficient time is allowed to elapse to allow binding ofthe APC with the disease organisms, and then, any excess fluid isdrained away by gravity or actively aspirated using a needle andsyringe.

[0123] Step 3 The light source is positioned behind the ear andactivated. The light source need not be very intense since the middleear cavity is small. Further, the fluence rate and total fluence dosemay be followed as discussed above.

Example 7

[0124] Transcutaneous PDT for Targeting Antibiotic Associated PseudoMembranous Colitis

[0125] In cases where Clostridium difficile causes pseudomembranouscolitis, the same scheme disclosed above for the treatment of H. pylorimay be applied. The difference is that the APC is targeted toward C.difficile and the ingested light source is activated in the colon ratherthan in the stomach. Alternatively, the photosensitive agent can beactivated with transcutaneously transmitted light from an external lightsource, or by ultrasonic energy produced by an ultrasonic transmitter.

Example 8

[0126] Transcutaneous PDT for Targeting Septic Shock Disease

[0127] A number of anti-endotoxin antibodies and peptides have beendeveloped and synthesized that can be bound to photosensitizers to formanti-endotoxin APCs. These APCs are injected, allowed to bind and thenactivated transcutaneously with light, or by using the intracorporeallight emitting devices disclosed in U.S. Pat. No. 5,702,432. Fortranscutaneous activation, an external light source is placed over amajor vessel, preferably an artery, but most preferably a vein where theblood flow is slower, to allow more time for APC activation.

Example 9

[0128] Liver Cancer Photodynamic Therapy by Transillumination

[0129] This Example uses the present invention for the treatment of anorgan infiltrated with tumor tissue. Reference is made to FIG. 9.Specifically, light 140 is administered by transillumination throughliver tissue 148 from an implanted light source 144 that is disposedexternal to the surface of liver 142, but within the patient's bodyunderneath the skin layer 18. In this embodiment, a patient is injectedintravenously with a photosensitizer agent ICG, conjugated to anantibody that is specific to vascular endothelial antigen (notseparately shown) on a tumor 146, so that the antibody binds with theantigen, but not to other tissue in the liver. The optimal dose of ICGwill be empirically determined, for example, via a dose escalationclinical trial as is so often performed to evaluate chemotherapeuticagents. One or more light source probes 144 are surgically implanted(e.g., endoscopically) adjacent to, but not invading parenchymal tissue148 of liver 142. After delaying a time sufficient to permit clearing ofthe photosensitizer conjugate from the non-target tissues, the lightsource(s) is(are) activated, irradiating the target tissue with light140 at a relatively low fluence rate, but administering a high totalfluence dose of light in the waveband from about 750 nm to about 850 nm.

[0130] The specific dose of photosensitizer conjugate administered tothe patient is that which will result in a concentration of active ICGin the blood of between about 0.01 μg/ml and about 100 μg/ml and morepreferably, between about 0.01 μg/ml and about 10 [μg/ml. It is wellwithin the skill of the ordinary skilled artisan to determine thespecific therapeutically effective dose using standard clinicalpractices and procedures. Similarly, a specific acceptable fluence rateand a total fluence dose may be empirically determined based upon theinformation provided in this disclosure.

[0131] Although the present invention has been described in connectionwith the preferred form of practicing it, those of ordinary skill in theart will understand that many modifications can be made thereto withinthe scope of the claims that follow. Accordingly, it is not intendedthat the scope of the invention in any way be limited by the abovedescription, but instead be determined entirely by reference to theclaims that follow.

What is claimed:
 1. A method for transcutaneously administering a therapy to a target tissue in a mammalian subject, comprising the steps of: (a) administering to the subject a therapeutically effective amount of a targeted compound that is activated by ultrasonic energy to render the therapy, where the targeted compound selectively binds to the target tissue, but not to a non-target tissue; and (b) irradiating at least a portion of the subject with ultrasonic energy at a wavelength that activates the targeted compound, causing the target tissue to be destroyed.
 2. The method of claim 1, wherein the step of irradiating is delayed for a period of time sufficient for any of the compound that has not bound to the target tissue to clear from adjacent non-target tissue of the mammalian subject, prior to the step of irradiating.
 3. The method of claim 1, wherein the target tissue is selected from the group consisting of a vascular endothelial tissue, an abnormal vascular wall of a tumor, a solid tumor in one of the head, the neck, the gastrointestinal tract, the liver, the breast, the prostate, and the lung, a nonsolid tumor, malignant cells in hematopoietic tissue, malignant cells in lymphoid tissue, lesions in a vascular system, diseased bone marrow, cells afflicted by an autoimmune disease and cells afflicted with an inflammatory disease.
 4. The method of claim 1, further comprising the step of providing an ultrasonic emitting source for emitting the ultrasonic energy used for the step of irradiating.
 5. The method claim 4, wherein the ultrasonic energy emitting source is disposed external to an intact skin layer.
 6. The method of claim 1, wherein said compound includes an energy-activated agent that is conjugated to a ligand.
 7. The method of claim 6, wherein the ligand is selected from the group consisting of a target-specific antibody, a target-specific peptide and a target-specific polymer.
 8. The method of claim 1, wherein the compound is selected from the group consisting of indocyanine green, methylene blue, toluidine blue, aminolevulinic acid, phthalocyanines, porphyrins, purpurins and texaphyrins.
 9. The method of claim 1, wherein the compound is selected from the group consisting of gallium-porphyrin complexes, protoporphyrin, hematoporphyrin, daunorubicin and adriamycin.
 10. The method of claim 1, wherein the step of irradiating is carried out for a time interval of from about 30 minutes to about 72 hours.
 11. The method of claim 1, wherein the ultrasonic energy used for the step of irradiating is at a frequency between about 5 kHz and more than about 300 MHz.
 12. The method of claim 1, wherein the ultrasonic energy used for the step of irradiating is at a frequency between about 20 kHz and more than about 100 MHz.
 13. The method of claim 1, wherein the targeted compound activated by ultrasonic energy comprises one of: (a) a targeted compound activated by ultrasonic energy; (b) a delivery system that delivers the targeted compound activated by ultrasonic energy to bind with the target tissue; or (c) a prodrug that produces a prodrug product activated by ultrasonic energy, wherein the prodrug product selectively binds to the target tissue.
 14. An ultrasonic energy-activated targeted delivery system that is selectively targeted at a target tissue, comprising: (a) an ultrasonic energy-activated agent that absorbs energy and destroys a target tissue to which it is bound; and (b) a ligand conjugated to the ultrasonic energy-activated agent, said ligand binding to a receptor on the target tissue with specificity, so that binding of the ligand to a non-target tissue is minimized.
 15. The ultrasonic energy-activated targeted delivery system of claim 14, wherein the energy-activated agent comprises a prodrug.
 16. The ultrasonic energy-activated targeted delivery system of claim 14, wherein the ligand comprises an antibody that binds to the receptor.
 17. The ultrasonic energy-activated targeted delivery system of claim 14, wherein said receptor is selected from the group consisting of a vascular endothelium antigen, an antigen that is specific for an abnormal vascular wall of a tumor and an antigen that is specific for a non-vascular tumor tissue.
 18. The ultrasonic energy-activated targeted delivery system of claim 14, wherein the ligand is selected from the group consisting of a target-specific antibody, a target-specific antibody fragment, a target-specific peptide and a target-specific polymer.
 19. The ultrasonic energy-activated targeted delivery system of claim 14, wherein the ligand and the receptor comprise a binding pair selected from the group consisting of a biotin-streptavidin, a chemokine-chemokine receptor, a growth factor-growth factor receptor and an antigen-antibody.
 20. The ultrasonic energy-activated targeted delivery system of claim 14, wherein the ligand is a target-specific antibody specific to an antigen selected from the group consisting of tumor surface antigen, tumor endothelial antigen, non-tumor endothelial antigen and tumor vessel wall antigen. 